| 秦灏,黄海杰,黄伟坚,张中润,夏益华,赵丽.腰果AoCDPK26基因的克隆、亚细胞定位及表达特性分析[J].中国南方果树,2024,53(6): |
| 腰果AoCDPK26基因的克隆、亚细胞定位及表达特性分析 |
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| 投稿时间：2024-06-26 修订日期：2024-08-29 |
| DOI： |
| 中文关键词: 关键词：腰果 AoCDPK26 亲缘关系 RT-PCR 亚细胞定位 |
| 英文关键词: |
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| 中文摘要: |
| CDPK多基因家族在植物的非生物胁迫中发挥着重要的作用，本研究旨在探究腰果(Anacardium occidentale L.)中AoCDPK26基因的功能和表达特性。本研究以腰果叶片为实验材料，通过RT-PCR扩增技术，获得了AoCDPK26基因序列，利用生物信息学分析方法对其理化性质、氨基酸的序列、蛋白质结构进行了初步分析预测，利用RT-qPCR技术对其在不同组织部位、低温和不同激素处理后的表达特性进行了测定。此外构建了pEGAD-35S::AoCDPK26-GFP 重组载体在烟草中对AoCDPK26基因进行了亚细胞定位。生物信息学分析表明，腰果AoCDPK26基因编码区序列含有一个1692bp的开放阅读框，编码564个氨基酸，系统进化树分析表明该序列与开心果具有较高的亲缘关系。RT-qPCR检测结果表明，AoCDPK26基因在腰果的根、茎、叶、花、果梨和果仁中具有表达，其中在腰果叶片组织中的表达量最高；在低温胁迫下，AoCDPK26基因在处理5d后的表达量达到最高，是对照表达量的38倍；用MeJA激素处理腰果叶片后，AoCDPK26基因在24h的表达量增幅最大。在烟草组织的亚细胞定位实验表明AoCDPK26定位于细胞膜和细胞核中。腰果AoCDPK26基因的可能参与低温胁迫和激素响应的调控。 |
| 英文摘要: |
| The CDPK multigenic family plays an important role in anti-abiotic stress of plants，the function and expression characteristics of AoCDPK26 gene in cashew (A. occidentale ) were investigated. The leaves of cashew were used as experimental materials, and the AoCDPK26 gene sequence was obtained by RT-PCR.The physicochemical properties, amino acid sequence and protein structure were preliminically predicted by bioinformatics analysis. Real-time fluorescence quantitative PCR was exployed to detect the expression of AoCDPK26 gene in different tissue of cashew seedling, different treatments with low temperature stress and multiple plant hormones. In addition, the recombinant vector PEGAD-35S:: AOCDPK26-GFP was constructed and subcellular localization of AoCDPK26 gene was carried out in tobacco.The results indicated that AoCDPK26 gene encoding a total length of 1692bp and 564 amino acids was a hydrophilic non-transmembrane protein without signal peptide, which had a high similarity with Pistacia vera. Subcellular localization demonstrated that AoCDPK26 was localized in cell membrane and nucleus. RT-qPCR showed that the expression of AoCDPK26 gene was the highest in the cashew leaves and the lowest in the cashew nuts. After 5 days of low temperature treatment, the expressionof AoCDPK26 gene peaked, reaching 38 times compared to the control. Notably, the greatest increase inAoCDPK26 gene expressionwas observed in cashew leaves treated with Methyl Jasmonate (MeJA) at the 24-hour mark.The AoCDPK26 gene of A. occidentale may be involved in the regulation of low temperature stress and hormone response. |
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